Organoids and Gene Editing Core

Washington University, Saint Louis MO

Investigators

Linked publications, trials & patents

Abstract

PROJECT SUMMARY OGEC The overarching mission of the WU-DDRCC is promoting collaborative, multidisciplinary research focused on interactions between host and environment in digestive disease. The reconfigured Organoid and Gene Editing Core (OGEC), and its predecessor, the Precision Animal Models and Organoids Core (PAMOC), supports this mission by offering human-derived and preclinical models along with the expertise to rigorously study digestive disorders. Guided by our External Advisory Board (EAB), member survey results and usage trends, OGEC has evolved to offer new services, techniques, and leveraged constructive collaborations with Institutional Cores. In the current cycle, we expanded the Organoid Component to meet member demand and devolved germ-free services to the new WUSTL Institutional gnotobiotic facility. New OGEC services introduced over the most recent funding period (2019-2024) include gene editing strategies (leveraged through Institutional core services), air-liquid-interface culture, hepanoid and hepatocellular tumoroid culture, multicellular culture including organ-on-a-chip, host-microbe studies, and acquisition of new equipment for imaging and monitoring organoid growth (Biotek Lionheart Fx). KEY ACCOMPLISHMENTS: Since 2019, the reconfigured OGEC has established 96 new human cell lines from 68 different patients including those with Crohn’s, UC, ileal pouches, IL-10R deficiency, and cancers of the stomach, liver and colon. We distributed 56 human cell lines to 11 DDRCC members. We established murine lines from 17 different genetic models and distributed these to 5 laboratories. We generated and distributed 160 L of QC tested L-WRN media as well as other reagents to 27 different members. Together, these services have provided member cost savings of $1.876M. The mouse genetics component completed 10 genome modification projects (increasingly with CRISPR/Cas9/targeting vector injections or electroporations) for 7 members; and provided 3 transgenic mouse strains from the Mouse Bank to 7 members. 28 Members used the Core publishing 36 papers of which 24 (67%) were collaborative involving 2 or more members citing DDRCC. Our SPECIFIC AIMS include Aim 1: Provide efficient access to organoids for research by offering useful human and mouse cell lines, discounted culture media, imaging equipment, and consultation services. Aim 2: Provide direct assistance for organoid experiments, including deriving gastrointestinal cell lines from mice and humans, editing organoid genomes, and providing wells/plates of high-quality organoids in 2D, 3D, and/or multicellular format. Aim 3: Assist members in generating gene-edited mice, including electroporating CRISPR/Cas9 reagents into zygotes. These services provide vital pre-clinical research tools and strengthen the relevance of our science to clinical practice. Aims 1 and 2 build on infrastructure and expertise from Dr. Ciorba and collaborators at WUSTL. Aims 2 and 3 leverage Institutional resources of the Genome Engineering & Stem Cell Center and the Mouse Genetics Core, with Dr. Miner offering guidance for members at each step.